Table 4. Summary of all results from Tcp12 reconstitution with different peptides (1–9, D-1, D-4) and the adenylation domain substrates Dpg and ATP. All peptides were presented bound to PCP6.
| Substrate | Yield
a
[%] |
|||
| Dpg extended peptide, PCP-bound b | Dpg extended peptide, hydrolysed c | PCP6-bound peptide d | Hydrolysed starting peptide e | |
| 1 | 52.8 ± 1.9 | 1.6 ± 0.1 | 40.8 ± 1.3 | 4.9 ± 0.9 |
| 2 | 64.2 ± 3.2 | 4.2 ± 1.1 | 16.8 ± 0.8 | 14.8 ± 1.8 |
| 3 | 76.4 ± 0.25 | 4.6 ± 0.81 | 7.7 ± 0.37 | 11.3 ± 0.7 |
| 4 | 73.9 ± 5.1 | 4.8 ± 0.6 | 11.0 ± 1.7 | 10.3 ± 5.6 |
| 5 | 47.8 ± 2.2 | 3.9 ± 0.4 | 45.4 ± 2.4 | 3.0 ± 0.3 |
| 6 | 61.3 ± 1.4 | 7.0 ± 1.3 | 19.7 ± 0.5 | 11.9 ± 0.3 |
| 7 | 8.6 ± 0.4 | 1.4 ± 0.1 | 80.5 ± 2.2 | 9.6 ± 2.5 |
| 8 | 48.5 ± 11.3 | 2.6 ± 1.1 | 41.1 ± 12.1 | 7.8 ± 0.9 |
| 9 | 64.3 ± 1.3 | 2.3 ± 0.2 | 27.9 ± 1.0 | 5.5 ± 0.3 |
| D-1 | 35.2 ± 8.1 | 8.6 ± 7.5 | 39.2 ± 10.8 | 17.1 ± 8.2 |
| D-4 | 55.9 ± 2.7 | 6.1 ± 0.2 | 27.3 ± 2.5 | 10.7 ± 0.2 |
aTotal yield of extended peptide is based on the percentage reduction of initial hexapeptide peak from initial starting material. The hydrolysed/PCP-bound fractions for each peptide length is determined by dividing the area for each peak by the sum of both peptide peaks.
bElongated product cleaved through the use of methylamine to cleave the PCP7-bound thioester.
cElongated product hydrolysed from PCP7 domain of Tcp12 construct during the course of the reaction.
dPCP6-bound peptide substrate cleaved with methylamine.
eStarting peptide hydrolysed from PCP6 during the course of the reaction.